体外培养的慢性淋巴细胞性白血病患者淋巴细胞的姊妹染色单体互换及细胞周期进程

人类血淋巴细胞中姊妹染色体互换(SCE)的发生频率是相当恒定的,也不因年龄和性别而有差异。但是无论是体内或是体外用一系列广为不同的致变剂—致癌剂处理都可使SCE大大增加,比起正常人来,一种罕见的遗传性疾病—Bloom氏综合征患者的SCE也增加许多倍。为研究恶性肿瘤或白血病患者SCE是否也有改变。本文作者对采自19例慢性淋巴细胞性白血病(CLL)患者及10例正常人的淋巴细胞进行了培养与比较。结果表明,接受治疗的慢性淋巴细胞性白血病患者,其     

急性白血病、慢性髓细胞白血病及骨髓增生异常综合征患者染色体分析

目的探讨染色体分析在急性白血病（AL）、慢性髓细胞白血病（CML）及骨髓增生异常综合征（MDS）诊断及预后判断中的价值。方法采用骨髓细胞短期培养法,应用G、R显带技术对238例AL、CML及MDS患者进行染色体分析。结果56.30%的患者有染色体异常,其中数目异常占6.30%,结构异常占39.08%,复杂异常占10.92%;各组患者中CML组异常检出率最高,占总病例的34.87%;AL组染色体畸变最为复杂,累及除3,4,5,10及X以外的所有染色体,11例患者出现了特异性染色体畸变,与患者预后相关;CML加速/急变期出现了附加染色体异常;核型异常的MDS患者均检出了8三体,7例为单纯8三体,2例RAEB-T患者均为复杂异常。结论染色体检查对于疾病诊断与鉴别、指导临床治疗、判断预后等具有重要意义。     

骨髓增生异常综合征与白血病细胞p15INK4B基因甲基化及机制研究

目的:探讨p15INK4B基因甲基化异常和血液系统肿瘤发病的关系及甲基化异常的机制。方法:采用RT-PCR、甲基化特异PCR、Western blot法检测20例骨髓增生异常综合征(MDS)、20例急性白血病患者(AL)、14例慢性粒细胞性白血病(CML)患者骨髓单个核细胞p15INK4B基因 mRNA和p15INK4B蛋白的表达、p15INK4B基因甲基化及甲基转移酶(DNMTs)的表达。结果:高危组MDS患者p15INK4B蛋白表达阳性率低于低危组MDS患者(10% vs 80%,P<0.01),p15INK4B基因甲基化阳性率较高(60% vs 10%,P<0.01)。20例AL有9例(45%)存在p15INK4B基因甲基化。10例CML慢性期(CML-CP)患者仅1例存在p15INK4B基因甲基化。4例CML急变期(CML-BP)患者均检测到p15INK4B基因部分甲基化。DNMT_3A、DNMT_3B表达在AL、高危组MDS和CML-BP患者均明显高于低危组MDS(P<0.05)。结论:p15INK4B基因甲基化在AL、高危组MDS和CML-BP患者发生率高于低危组MDS,伴有甲基转移酶DNMT_3A、DNMT_3B表达较高。p15INK4B基因甲基化可能参与MDS和AL的发病机制并与MDS 及CML的预后有关。     

骨髓增生异常/骨髓增殖性肿瘤1例诊治分析

了解骨髓增生异常/骨髓增殖性肿瘤(myelodysplastic/myeloproliferative neoplasms,MDS/MPN)的临床类型、病理特征、基因突变,对其预后进行评估.收集1例MDS/MPN病例及相关参考文献,观察其临床表现、临床病理学特征、基因突变及最终临床分型,判断治疗疗效,并进行预后评价.MDS/MPN同时具有MDS和MPN的临床及主要血液学特点,目前分为慢性粒单细胞白血病、不典型慢性粒细胞白血病,BCR-ABL1阴性(atypical chronic myeloid leukemia,aCML)、幼年型粒单细胞白血病(juvenile myelomonocytic leukemia,JMML)、伴环形铁粒幼细胞和血小板增多的骨髓增生异常/骨髓增殖性肿瘤(MDS/MPN with ring sideroblasts and thrombocytosis,MDS/MPN-RS-T)、MDS/MPN,无法分类(MDS/MPN-unclassifiable,MDS/MPN-U)等5种类型.各型有其不同诊断标准,而上述不同疾病之间出现转化,国内未见报道.保定市第一医院诊治1例最初诊断不明确的慢性髓系恶性肿瘤,随着疾病的演变而最终确诊为慢性粒单细胞白血病.通过本病例表明,在不同类型MDS/MPN之间可能会存在类型转换.     

骨髓增生异常综合征患者的asxli基因变异分析

目的探讨骨髓增生异常综合征(myelodysplastic syndromes,MDS)患者ASXL1基因变异的发生情况及其与其他基因变异和部分临床参数之间的相关性。方法采用PCR扩增产物直接测序法检测149例MDS患者ASXU、U2AF1、SF3B1、DNMT3A、TET2、IDH1/2、NPM1、FLT3-ITD.C-KIT等基因的变异情况。结果在149例患者中,ASXU基因变异的检出率为24.8%(37/149),变异率>5%的基因分别是U2AF1(22.8%)、TET2(11.4%)、DNMT3A(9.4%)、NPM1(8.1%).SF3B1(6.0%)。ASXL1变异最常见的共存变异基因为U2AF1(27.0%,10/37)及TET2(18.9%,7/37)。ASXL1变异组与野生组患者在中位年龄、MDS亚型、染色体核型、外周白细胞、血红蛋白、血小板水平及骨髓原始细胞计数等方面的差异均无统计学意义(P>0.05)o对29例ASXL1变异患者进行了有效的随访,其中11例进展为急性髓系白血病(acute myeloid leukemia,AML),白血病转化率为37.9%。在92例野生型患者中,13例进展为AML,白血病转化率为14.1%。ASXL1变异组白血病转化率明显高于野生组,差异有统计学意义(PV 0.01)。结论ASXL1变异在MDS中有较高的发生率,并常与U2AF1及TET2基因变异共存,伴有该变异的患者具有更高的白血病转化率。     

血清铁蛋白与骨髓增生异常综合征危险度分层和预后的关系分析

目的 探讨血清铁蛋白水平在骨髓增生异常综合征(MDS)患者中危险度分层及预后评估的临床意义.方法 对41例初诊MDS患者和30名健康对照者采用电化学发光免疫法测量血清铁蛋白(SF),观察MDS患者SF水平及与国际预后评分系统(IPSS)的关系.对MDS患者进行随访,根据铁蛋白水平和IPSS评分的数值绘制ROC曲线,并将患者分组分为相对低危组(SF＜ 577ng/mL)和相对高危组(SF≥577ng/mL),比较两组间白血病转化率和生存时间.结果 MDS组患者铁蛋白水平明显高于健康对照组(P＜0.05),相对低危组与相对高危组之间铁蛋白水平差异有统计学意义(P＜0.05),随访的39位患者中共12人转化为急性白血病,两组转化率分别为10.5％和45.5％,向白血病转化的中位时间分别为18.5(4 ～46)个月和30.0(6 ～48)个月,差异有统计学意义(P＜0.05),两组间生存时间差异无统计学意义(P＞0.05).结论 铁蛋白在MDS患者中高于正常水平,初诊未输血的患者存在铁过载,高危类型MDS患者的SF水平较高,SF水平与其不良预后有一定的相关性.     

血清乳酸脱氢酶、铁蛋白和血小板生成素在急性白血病与骨髓异常增生综合征鉴别诊断中的应用

目的:研究血清乳酸脱氢酶(Lactate dehydro-genase,LDH)、铁蛋白(Serum ferritin,SF)和血小板生成素(Thrombopoietin,TPO)对急性白血病(Acute leukemia,AL)和骨髓异常增生综合征(Myelodysplastic syndrome,MDS)鉴别诊断价值.方法:选取2016年9月至2018年9月我院收治的AL患者71例(AL组)和MDS患者52例(MDS组),测定血清LDH、SF和TPO水平,并采用ROC曲线分析LDH、SF、TPO及三者联合检测对两种疾病的鉴别诊断价值.结果:AL组血清LDH及TPO水平明显高于MDS组,血清SF水平明显低于MDS组(P<0.05);三者与AL均密切相关(P<0.05),对AL和MDS鉴别诊断AUC分别为0.684、0.814和0.922,且三者联合诊断AUC达0.963,较单独应用明显升高(P<0.05).结论:血清LDH、SF和TPO在AL和MDS患者中表达水平存在显著差异,对临床鉴别诊断具有良好参考意义.     

骨髓增生异常综合征血清β2—m临床观察及意义

本文对31例骨髓增生异常综合征（MDS）患者进行血清β_2-微球蛋白（β_2-m）检测,为了解血清β_2-m水平在MDS中的变化,并对31例MDS患者不同病期进行了动态观察,现将结果报告如下。 材料和方法 一、对象: （一）正常对照组:其30例（男20,女10）,年龄19～58岁,均为本院健康职工,无肾、肝、血液等相关疾病。 （二）观察组:MDS共31例（男14,女17）,年龄14～65岁,其中MDS（RA）期9例;MDS（RAS）期4例;MDS（RAEB）期7例;MDS（RAEB-T）期6例;转化型白血病期5例。所有病例均经临床和实验室证实。按全国血液病学术会议拟定的MDS诊断和分型标准。 二、方法: （一）采用放射免疫分析法（RIA）,β_2-m试剂盒     

获得性21三体髓系恶性血液病的生物学特征及预后意义

目的：系统分析21三体髓系恶性血液病临床特征及预后意义。方法：应用常规细胞遗传学技术对536例急性髓系白血病（AML）、145例骨髓增生异常综合征（MDS）进行检测，随访疗效及生存状况。结果：18例21三体患者在AML和MDS的发病率分别为1．68％和6．21％。作为孤立异常分别为0．18％及2．74％。伴有结核病、银屑病及毒物接触史患者（38．89％）比非21三体患者（8．45％）显著增多（P＜0．01）。AML以M2a多见。MDS转白血病均为M4a。55．56％AML获得完全缓解，中位生存期10个月。5例M2a型中位生存期仅为3个月，与染色体核型正常患者相比显著缩短（P＜0．05）。结论：原发或孤立出现的21三体AML通常与M2a型相关，且多有其他疾病史或毒物接触史，预后极差。MDS转白类型常为M4a，预后不良。     

造血干细胞移植治疗骨髓增生异常综合征20例疗效分析

目的评价造血干细胞移植（HSCT）治疗骨髓增生异常综合征（MDS）患者的疗效，探讨MDS患者接受HSCT治疗的适应证和时机。方法1993年11月～2007年4月对20例MDS及MDS转急性髓系白血病（AML）患者（男性12例,女性8例,中位年龄39岁）行HSCT治疗。其中18例接受同胞供者异基因外周血干细胞移植（allo—PBSCT）；1例为同基因骨髓移植（Svn—BMT）；1例为无关脐带血移植（CBT），＋25d时移植失败行自体骨髓移植。预处理主要采用修改的Bu／Cv方案。结果3年总生存率（SO）及3年无病生存率均为53.3％±12％；3年复发率（RR）10．8％±7％，移植相关死亡率（TRM）42．6％±12％。截止随访日期，存活11例，中位生存时间16．5（2.0～112）个月。结论HSCT是治疗MDS的有效方法,如有HLA匹配的同胞供者,HSCT可作为MDS患者的一线治疗。     

DNA损伤修复与白血病存活关系的研究

本文以自发和ＭＭＣ诱发的ＳＣＥ值为指标，分析了临床完全缓解后白血病患者ＳＣＥ值的改变。结果显示，临床完全缓解８例患者自发ＳＣＥ值与正常对照无显著差异（Ｐ＞０．０５），ＭＭＣ诱发的ＳＣＥ值两组间差异极显著（Ｐ＜０．０１）。说明患者的染色体仍有潜在的不稳定性，应继续巩固治疗。临床完全缓解４、５年以上者，自发和诱发ＳＣＥ值与正常对照均无差异（Ｐ＞０．５），提示临床缓解时间长，遗传物质相对稳定。     

Sister chromatid exchanges in leukemic patients

Sister chromatid exchange (SCE) was studied in PHA-stimulated peripheral blood lymphocytes from 36 newly diagnosed and untreated leukemic patients: 16 with acute lymphoblastic leukemia (ALL), 10 with acute nonlymphocytic leukemia (ANLL), and 10 with chronic myelocytic leukemia (CML). The metaphases analyzed show no chromosomal abnormalities. The mean SCE frequency (mean +/- SE) for each group of patients was: 6.8 +/- 0.4, 6.6 +/- 0.3, and 7.0 +/- 0.6 per mitosis, respectively, which was significantly lower than the mean SCE score for 30 controls (8.7 +/- 0.2). No differences in SCE score among ALL, ANLL, and CML and a similar SCE frequency by chromosome number and group allowed consolidation of all the cases into a single group of 36 leukemic patients (6.8 +/- 0.3). When the frequency of SCE was compared by chromosome number and group between the leukemic patients with the control group, a significant decrease in SCE frequency was observed due to a low SCE score in almost all the complements, except chromosome #1. It is suggested that the low SCE rate is related to the leukemic process itself.     

Cell-cycle progression rates and sister chromatid exchange frequencies in the bone marrow of patients with myelodysplastic syndrome and acute myeloid leukemia.

The proliferation characteristics of leukemic cells may be a determining factor in disease course and response to therapy. The present study compares the rate of cell-cycle progression in the bone marrow of 16 hematologically normal subjects, 19 patients with acute myeloid leukemia (AML), and 23 patients with myelodysplastic syndrome (MDS). The frequency of sister chromatid exchanges (SCE) in bone marrow cells is also compared. MDS and AML patients showed a reduction in the rate of cell-cycle progression compared with normal subjects. Patients with 'high risk' MDS (RAEB/RAEB-t) did not differ significantly from patients with AML but had a significantly slower rate of cell-cycle progression than patients with 'low-risk' MDS (PASA/RA). There was no correlation between the rate of cell-cycle progression and clonal karyotype status or the percentage of blast cells in either MDS or AML. There were no significant differences in SCE frequency between normal subjects and MDS or AML patients.     

Increased spontaneous and mitomycin C-induced sister chromatid exchanges in patients with cancer of the cervix uteri, with special reference to stage of cancer

The frequencies of spontaneous and mitomycin C (MMC)-induced sister chromatid exchange (SCE) were examined in 35 patients with cancer of the cervix uteri (stage 0, eight cases; stage I, nine cases; stage II, nine cases, and stage III, nine cases) before they had undergone cancer treatment, as well as in seven patients with uterine myoma and 18 healthy women as controls. The frequency of SCE was analyzed in reference to the stage of cancer in the cancer group and in reference to chromosome group in the cancer and normal groups. The frequencies of spontaneous and MMC-induced SCE in the cancer group were 10.0 +/- 1.8 and 20.7 +/- 2.6, respectively, and both were significantly higher than in the myoma (8.1 +/- 0.8 and 17.6 +/- 1.8) and normal (7.6 +/- 0.8 and 17.6 +/- 2.3) groups. Furthermore, the frequency of SCE in the cancer group increased with cancer stage. All chromosome groups contributed equally to the increase in SCE in the cancer group. These results indicate that an increase in the frequency of SCE in patients with cervical cancer is related to the presence of cancer, but is not related to a predisposition to cancer.     

Chromosome abnormalities, sister chromatid exchanges, and cell cycle analysis in phytohemagglutinin-stimulated adult T cell leukemia lymphocytes

Chromosome abnormalities, sister chromatid exchanges (SCE), and cell cycle kinetics were studied in phytohemagglutinin-stimulated lymphocytes from 8 adult T cell leukemia (ATL) patients. In all these cases, chromosome abnormalities were observed in 5-day PHA-stimulated cultures. Four cases had characteristic marker chromosomes; two were due to a balanced translocation, t(9;21), and two to a simple deletion, 5p-. The other four cases, however, had rather complicated chromosome abnormalities, e.g., 1q+, 2q+, 5q+, 6q-, 14q+, 10p-. When chromosome abnormalities were analyzed in previously reported cases, the abnormalities were mostly distributed among chromosomes #1, #2, #5, #6, #14, and #21. These findings suggest that the abnormalities involving #1, #2, #5, #6, #14, and #21 are intimately related to ATL. The SCE frequency was in the normal range in ATL cells. Cell cycle analysis revealed that the duration of two cell cycles in cells labeled with bromodeoxyuridine (BrdU) required approximately 80 hr in ATL cells, whereas the time of two cell cycles in normal cells is 40 hr. These findings indicate that the ATL cell cycle time is about 40 hr, about double that of normal cells (20 hr), in PHA-stimulated cultures. ATL has been known to be a mature T cell leukemia and to respond poorly to chemotherapy. The latter may be due to the elongated cell cycle or to the mature characteristics of the leukemic cells. The association of ATL with cutaneous T cell lymphoma is also discussed.     

伴复杂核型异常的髓系恶性血液病中17号染色体异常分析

目的 研究伴复杂核型异常(complex chromosomal abnormalities,CCAs)的髓系恶性血液病中17号染色体的异常特征.方法 经R显带常规细胞遗传学分析显示CCAs的73例髓系恶性血液病,包括21例急性髓系白血病(acute myeloid leukemia,AML)、36例慢性髓系白血病(chronic myeloid leukemia,CML)、16例骨髓增生异常综合征(myelodysplastic syndrome,MDS),并进一步多重荧光原位杂交分析.结果 73例伴CCAs的髓系恶性血液病中,17号染色体异常最常见,占46.5%(34/73),其中AML12例,CML13例,MDS9例,9例CML慢性期患者均未见17号染色体异常.结构异常较多见,总发生率为43.8%(32/73);AML、CML、MDS3组发生率分别为52.4%(11/21)、33.3%(12/36)、56.3%(9/16);所有病例中发生数目异常共15.1%(11/73),三组发生率分别为25.0%(3/12)、38.5%(5/13)、33.3%(3/9),11例数目异常均为-17.有9例同时出现数目异常和结构异常.伴有17号染色体的结构异常中,以非平衡易位多见,3组分别为16、15、8个;平衡易位2个,分别为发生于AML中的t(15;17)及发生于CML中的t(15;17;22).17号染色体结构易位的对手染色体多变,包括了除5号、6号和22号外的所有染色体.结构易位频率最高的对手染色体是15号,占8.2%(6/73);其次为2号,占5.4%(4/73).6例存在17号与15号易位的病例中5例为急性早幼粒白血病,1例为CML急变期.结论 伴CCAs的髓系恶性血液病中17号染色体异常发生率高,以结构异常为主.所有的数目异常均为-17;结构异常以非平衡易位多见.     

Transformation of the 5q- syndrome to acute lymphoblastic leukemia: a report of two cases and review of the literature

Myelodysplastic syndrome (MDS) with an isolated deletion of the long arm of chromosome 5 (5q- syndrome) is a distinct subtype of MDS with an indolent course that rarely transforms to acute leukemia. Deletion of the long arm of chromosome 5 has also been reported in rare cases of de novo B-lymphoblastic leukemia. We present two cases of 5q- syndrome with a similar and unusual course of transformation to lymphoblastic leukemia while on Lenalidomide. These two patients achieved an initial response; however, later acquired a second cytogenetic abnormality, became refractory to treatment and evolved into acute leukemia. At the time of transformation, both patients had recurrence of the 5q- abnormality. Review of the literature and the mechanisms of transformation of the 5q-syndrome into an acute leukemia are discussed. Although the relationship between the events in our cases remains unclear, the intriguing similarity between the two cases raises a question whether immune modulators can alter the natural course of MDS. To our knowledge, no similar cases were previously reported in the literature.     

p53 expression in myeloid cells of myelodysplastic syndromes. Association with evolution of overt leukemia.

To assess p53 expression in the hematopoietic cells of the bone marrow in premalignant as well as malignant conditions, we examined immunohistochemically bone marrow biopsies from patients with myelodysplastic syndromes (MDS, n = 51), acute myeloid leukemia (n = 42) and as a nonneoplastic condition, aplastic anemia (n = 20) and samples from individuals who had no hematological disorder (control, n = 12). Nuclear accumulation of p53 protein was found in seven of 51 patients with MDS (14%) and two of 42 acute myeloid leukemia patients (5%), whereas patients with aplastic anemia and control subjects were uniformly negative for p53 protein. In the bone marrow of patient with MDS, p53-positive cells constituted about 5 to 30% of the total bone marrow cells. Two-color immunohistochemical analysis revealed that the p53-positive cells were also positive for the myeloid cell marker. Half of the MDS cases that evolved to overt leukemia (seven of 14) exhibited positive p53 reaction in the bone marrow at the time of initial diagnosis. This frequency (50%) was significantly higher than that in de novo acute myeloid leukemia cases. All of the seven MDS cases that exhibited p53 expression at the time of initial diagnosis developed overt leukemia later, and p53 expression was maintained throughout the progression of MDS. The results suggest that p53 mutations that occur in the myeloid cells in MDS may confer a growth advantage to these cells resulting in the progression to overt leukemia. Thus, immunohistochemical examination for p53 is very useful for predicting the evolution to overt leukemia from MDS.     

Clinical significance of Y chromosome loss in hematologic disease

We have studied 215 male patients (aged 45-97 years) whose sole cytogenetic abnormality was clonal loss of the Y chromosome in metaphase cells from unstimulated cultures. The patients comprised a control group with no evidence of hematologic disease and four disease case groups: 1) myelodysplastic syndrome (MDS), refractory anemia, refractory anemia with excess blasts (RAEB), RAEB in transformation, and chronic myelomonocytic leukemia; 2) acute myelogenous leukemia; 3) myeloproliferative disorder (MPD), chronic granulocytic leukemia, and polycythemia vera; and 4) B-cell lymphoma/leukemia. The frequency of cells with Y loss increased with age and was significantly greater in cases than in controls, but it was not correlated with survival or with prior therapy. The frequency of cases with a -Y clone was 6.3% of male karyotypes and represented 16.4% of all abnormal male cytogenetic reports. Much of the difference between cases and controls appears to be accounted for by a greater frequency of cases with > 75% Y loss. A value of 81% chromosome Y loss maximized the combined sensitivity (28%) and specificity (100%) for predicting disease status, but a 75% cutoff provided the best estimate of disease risk. Even in older males, if > 75% of metaphase cells are 45,X,-Y, they probably represent a disease-associated clonal population, and it is possible that the critical genetic change is not visible through the microscope. This observation is true for MDS, MPD, B-cell disease, and especially acute myelogenous leukemia. The prognostic association of Y chromosome loss for survival appears to be neutral or favorable. Genes Chromosomes Cancer 27:11-16, 2000.     

Genomic Copy Number Variations in the Myelodysplastic Syndrome and Acute Myeloid Leukemia Patients with del(5q) and/or -7/del(7q)

The most common chromosomal abnormalities in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML) are -5/del(5q) and -7/del(7q). When -5/del(5q) and -7/del(7q) coexist in patients, a poor prognosis is typically associated. Given that -5/del(5q) and/or -7/del(7q) often are accompanied with additional recurrent chromosomal alterations, genetic change(s) on the accompanying chromosome(s) other than chromosomes 5 and 7 may be important factor(s) affecting leukemogenesis and disease prognosis. Using an integrated analysis of karyotype, FISH and array CGH results in this study, we evaluated the smallest region of overlap (SRO) of chromosomes 5 and 7 as well as copy number alterations (CNAs) on the other chromosomes. Moreover, the relationship between the CNAs and del(5q) and -7/del(7q) was investigated by categorizing the cases into three groups based on the abnormalities of chromosomes 5 and 7 [group I: cases only with del(5q), group II: cases only with -7/del(7q) and group III: concurrent del(5q) and del(7q) cases]. The overlapping SRO of chromosome 5 from groups I and III was 5q31.1-33.1 and of chromosome 7 from groups II and III was 7q31.31-q36.1. A total of 318 CNAs were observed; ~ 78.3% of them were identified on chromosomes other than chromosomes 5 and 7, which were defined as ''other CNAs''. Group III was a distinctive group carrying the most high number (HN) CNAs, cryptic CNAs and ''other CNAs''. The loss of TP53 was highly associated with del(5q). The loss of ETV6 was specifically associated with group III. These CNAs or genes may play a secondary role in disease progression and should be further evaluated for their clinical significance and influence on therapeutic approaches in patients with MDS/AML carrying del(5q) and/or -7/del(7q) in large-scale, patient population study.